Analytical and experimental study of reentrant stream crossed-field amplifiers Final report

Computer simulation and noise measurements of reentrant stream crossed-field amplifier

Direct imaging of RecA nucleation and growth on single molecules of SSB-coated ssDNA.

Escherichia coli RecA is the defining member of a ubiquitous class of DNA strand-exchange proteins that are essential for homologous recombination, a pathway that maintains genomic integrity by repairing broken DNA. To function, filaments of RecA must nucleate and grow on single-stranded DNA (ssDNA) in direct competition with ssDNA-binding protein (SSB), which rapidly binds and continuously sequesters ssDNA, kinetically blocking RecA assembly. This dynamic self-assembly on a DNA lattice, in competition with another protein, is unique for the RecA family compared to other filament-forming proteins such as actin and tubulin. The complexity of this process has hindered our understanding of RecA filament assembly because ensemble measurements cannot reliably distinguish between the nucleation and growth phases, despite extensive and diverse attempts. Previous single-molecule assays have measured the nucleation and growth of RecA--and its eukaryotic homologue RAD51--on naked double-stranded DNA and ssDNA; however, the template for RecA self-assembly in vivo is SSB-coated ssDNA. Using single-molecule microscopy, here we directly visualize RecA filament assembly on single molecules of SSB-coated ssDNA, simultaneously measuring nucleation and growth. We establish that a dimer of RecA is required for nucleation, followed by growth of the filament through monomer addition, consistent with the finding that nucleation, but not growth, is modulated by nucleotide and magnesium ion cofactors. Filament growth is bidirectional, albeit faster in the 5'→3' direction. Both nucleation and growth are repressed at physiological conditions, highlighting the essential role of recombination mediators in potentiating assembly in vivo. We define a two-step kinetic mechanism in which RecA nucleates on transiently exposed ssDNA during SSB sliding and/or partial dissociation (DNA unwrapping) and then the RecA filament grows. We further demonstrate that the recombination mediator protein pair, RecOR (RecO and RecR), accelerates both RecA nucleation and filament growth, and that the introduction of RecF further stimulates RecA nucleation

Migration to the US among rural Puerto Ricans who inject drugs: influential factors, sources of support, and challenges for harm reduction interventions

Background: While PWID of Puerto Rican origin have been migrating to the US for decades, the range of factors influencing their migration to the US and the resources they draw on to do so are not well understood. This is particularly true for rural Puerto Rican PWID, and the present study is the first empirical research to document migration patterns among this population. The specificities of their migration raise important challenges that need to be documented in order to implement more effective harm reduction policies at home (Puerto Rico) and abroad (US). Methods: This paper draws from data obtained employing a modified NHBS survey which was administered to (N =296) PWID in four rural municipalities of Puerto Rico with participants 18 years or older. The primary dependent variables for this paper are the number of times a person has lived in the continental US, and if they are planning on moving to the continental US in the future. Results: Findings suggest that 65% of the sample reported ever lived in the US and that 49% are planning on moving in the future. The number of times living in the US is associated with higher education and older age, but not with self-reported positive HIV or HCV statuses. Planning to move to the US is associated with knowing PWID who have moved or plan to move, negatively associated with age, and is not associated with HIV or HCV status. Around one-third of those that lived in the US reported having some sort of support, with the majority receiving support from family sources. No participant received help to enter HIV/HCV treatment. Conclusions: A multi-region approach to prevention is required to make a dent in curbing HIV/HCV transmission in this population. Understanding PWID migration patterns, risk behaviors, and health care needs in the US is now more important than ever as natural disasters prompted by human-made climate change will only increase in the future, raising demands not only for service providers but also harm reduction policies to cope with an increasing influx of “climate refugees” as PWID move across national borders

Measurements of the effect of horizontal variability of atmospheric backscatter on dial measurements

The horizontal variability of atmospheric backscatter may have a substantial effect on how Differential Absorption Lidar (DIAL) data must be taken and analyzed. To minimize errors, lidar pulse pairs are taken with time separations which are short compared to the time scales associated with variations in atmospheric backscatter. To assess the atmospheric variability for time scales which are long compared to the lidar pulse repetition rate, the variance of the lidar return signal in a given channel can be computed. The variances of the on-line, off-line, and ration of the on-line to off-line signals at given altitudes obtained with the dual solid-state Alexandrite laser system were calculated. These evaluations were made for both down-looking aircraft and up-looking ground-based lidar data. Data were taken with 200 microsecond separation between on-line and off-line laser pulses, 30 m altitude resolution, 5 Hz repetition rate, and the signal were normalized for outgoing laser energy

Real-time atmospheric absorption spectra for in-flight tuning of an airborne dial system

Real-time measurements of atmospheric absorption spectra are displayed and used to precisely calibrate and fix the frequency of an Alexandrite laser to specific oxygen absorption features for airborne Differential Absorption Lidar (DIAL) measurements of atmospheric pressure and temperature. The DIAL system used contains two narrowband tunable Alexandrite lasers: one is electronically scanned to tune to oxygen absorption features for on-line signals while the second is used to obtain off-line (nonabsorbed) atmospheric return signals. The lidar operator may select the number of shots to be averaged, the altitude, and altitude interval over which the signals are averaged using single key stroke commands. The operator also determines exactly which oxygen absorption lines are scanned by comparing the line spacings and relative strengths with known line parameters, thus calibrating the laser wavelength readout. The system was used successfully to measure the atmospheric pressure profile on the first flights of this lidar, November 20, and December 9, 1985, aboard the NASA Wallops Electra aircraft

Airborne Lidar measurements of the atmospheric pressure profile with tunable Alexandrite lasers

The first remote measurements of the atmospheric pressure profile made from an airborne platform are described. The measurements utilize a differential absorption lidar and tunable solid state Alexandrite lasers. The pressure measurement technique uses a high resolution oxygen A band where the absorption is highly pressure sensitive due to collision broadening. Absorption troughs and regions of minimum absorption were used between pairs of stongly absorption lines for these measurements. The trough technique allows the measurement to be greatly desensitized to the effects of laser frequency instabilities. The lidar system was set up to measure pressure with the on-line laser tuned to the absorption trough at 13147.3/cm and with the reference laser tuned to a nonabsorbing frequency near 13170.0/cm. The lidar signal returns were sampled with a 200 range gate (30 vertical resoltion) and averaged over 100 shots

Cyber War, Cybered Conflict, and the Maritime Domain

Those who dismiss cyber war as mere hype or as driven by potential profits dismiss much too quickly growing evidence of the importance of cyber operations—to which the Navy may be uniquely qualified to adapt

Transformation of Epichloë typhina by electroporation of conidia

<p>Abstract</p> <p>Background</p> <p>Choke, caused by the endophytic fungus <it>Epichloë typhina</it>, is an important disease affecting orchardgrass (<it>Dactylis glomerata </it>L.) seed production in the Willamette Valley. Little is known concerning the conditions necessary for successful infection of orchardgrass by <it>E. typhina</it>. Detection of <it>E. typhina </it>in plants early in the disease cycle can be difficult due to the sparse distribution of hyphae in the plant. Therefore, a sensitive method to detect fungal infection in plants would provide an invaluable tool for elucidating the conditions for establishment of infection in orchardgrass. Utilization of a marker gene, such as the green fluorescent protein (GFP), transformed into <it>Epichloë </it>will facilitate characterization of the initial stages of infection and establishment of the fungus in plants.</p> <p>Findings</p> <p>We have developed a rapid, efficient, and reproducible transformation method using electroporation of germinating <it>Epichloë </it>conidia isolated from infected plants.</p> <p>Conclusions</p> <p>The GFP labelled <it>E. typhina </it>provides a valuable molecular tool to researchers studying conditions and mechanisms involved in the establishment of choke disease in orchardgrass.</p

Migration to the US among rural Puerto Ricans who inject drugs: influential factors, sources of support, and challenges for harm reduction interventions

Background: While PWID of Puerto Rican origin have been migrating to the US for decades, the range of factors influencing their migration to the US and the resources they draw on to do so are not well understood. This is particularly true for rural Puerto Rican PWID, and the present study is the first empirical research to document migration patterns among this population. The specificities of their migration raise important challenges that need to be documented in order to implement more effective harm reduction policies at home (Puerto Rico) and abroad (US). Methods: This paper draws from data obtained employing a modified NHBS survey which was administered to (N =296) PWID in four rural municipalities of Puerto Rico with participants 18 years or older. The primary dependent variables for this paper are the number of times a person has lived in the continental US, and if they are planning on moving to the continental US in the future. Results: Findings suggest that 65% of the sample reported ever lived in the US and that 49% are planning on moving in the future. The number of times living in the US is associated with higher education and older age, but not with self-reported positive HIV or HCV statuses. Planning to move to the US is associated with knowing PWID who have moved or plan to move, negatively associated with age, and is not associated with HIV or HCV status. Around one-third of those that lived in the US reported having some sort of support, with the majority receiving support from family sources. No participant received help to enter HIV/HCV treatment. Conclusions: A multi-region approach to prevention is required to make a dent in curbing HIV/HCV transmission in this population. Understanding PWID migration patterns, risk behaviors, and health care needs in the US is now more important than ever as natural disasters prompted by human-made climate change will only increase in the future, raising demands not only for service providers but also harm reduction policies to cope with an increasing influx of “climate refugees” as PWID move across national borders

What's the Difference? Measures of Racial Disparity in Rates of a Sexually Transmitted Disease

Background: Elimination of racial disparities in health is a national health priority, yet little attention has been devoted to the choice of measures used to quantify disparity. Community-level risk factors for racial disparity in STDs are largely unstudied. Goal: To determine whether ten county-level demographic variables were associated with black-white disparity in gonorrhea incidence rates in North Carolina and to investigate how the association between the variables and racial disparity varied depending upon the measure of disparity used [incidence rate ratio (RR) vs. incidence rate difference (RD)]. Methods: We examined the relationships between the demographic variables and 5-year county average black-white RR and RD in gonorrhea in NC using simple linear regression, scatter plots, and Pearson's correlations. Results: All variables except sex ratio were more strongly correlated with RD than with RR. RD was strongly positively correlated with the incidence rate of gonorrhea among blacks (>0.99) and RR was less so (0.30). Conclusions: Several county-level variables are associated with additive racial disparity in gonorrhea in NC. This is likely due to the fact that RD is highly correlated with gonorrhea rates in the black population, and correlation of the variables with RD essentially reflects correlation with gonorrhea rates in the black population. Public health interventions aimed at reducing racial disparity in gonorrhea in NC should primarily aim to reduce absolute disparities.Master of Public Healt